EP 2812441 A1 20141217 - NUCLEIC ACID LIGATION METHOD
Title (en)
NUCLEIC ACID LIGATION METHOD
Title (de)
NUCLEINSÄURELIGATIONSVERFAHREN
Title (fr)
PROCÉDÉ DE LIGATURE D'ACIDES NUCLÉIQUES
Publication
Application
Priority
- US 201261595482 P 20120206
- US 2013024965 W 20130206
Abstract (en)
[origin: US2013203635A1] Methods and kits for covalently joining a 3' nucleic acid fragment having a 5'-hydroxyl terminus to a 5' nucleic acid fragment having a 3'-phosphate terminus are disclosed. The methods include the step of contacting the 3'-phosphate terminus of a first nucleic acid molecule and the 5'-hydroxyl terminus of a second nucleic acid molecule with an isolated 2',3'-cyclic phosphate RNA ligase (RtcB) and a purine triphosphate in the presence of manganese (II) ion, whereby the 3'-phosphate terminus of the first nucleic acid molecule and the 5'-hydroxyl terminus of the second nucleic acid molecule are covalently joined. Although the purine triphosphate used is generally GTP or dGTP, if the method is performed in the presence of an Archease, any purine triphosphate may be used. Accordingly, the disclosed kits include isolated RtcB, along with a purine triphosphate and/or an isolated Archease. Such methods and kits can be used to tag or ligate DNA or RNA on its 3'-phosphate terminus, as long as the terminal residue at the 3'-phosphate terminus is an RNA nucleotide.
IPC 8 full level
C12P 19/34 (2006.01); C07H 21/02 (2006.01); C07H 21/04 (2006.01); C12N 15/10 (2006.01); C12Q 1/68 (2006.01)
CPC (source: EP US)
C12N 9/00 (2013.01 - EP US); C12N 9/93 (2013.01 - EP US); C12N 15/1093 (2013.01 - EP US); C12P 19/34 (2013.01 - EP US)
Citation (search report)
See references of WO 2013119690A1
Citation (examination)
WO 2013160291 A2 20131031 - IMBA INST MOLEKULARE BIOTECH [AT]
Designated contracting state (EPC)
AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
Designated extension state (EPC)
BA ME
DOCDB simple family (publication)
US 2013203635 A1 20130808; EP 2812441 A1 20141217; WO 2013119690 A1 20130815
DOCDB simple family (application)
US 201313760920 A 20130206; EP 13706339 A 20130206; US 2013024965 W 20130206